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Molecular Mechanisms of Coupling to Voltage Sensors in Voltage-Evoked Cellular Signals

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Molecular Mechanisms of Coupling to Voltage Sensors in Voltage-Evoked Cellular Signals No. 3] Proc. Jpn. Acad., Ser. B 95 (2019) 111 Review Molecular mechanisms of coupling to voltage sensors in voltage-evoked cellular signals † By Yasushi OKAMURA*1,*2, and Yoshifumi OKOCHI*1 (Communicated by Masanori OTSUKA, M.J.A.) Abstract: The voltage sensor domain (VSD) has long been studied as a unique domain intrinsic to voltage-gated ion channels (VGICs). Within VGICs, the VSD is tightly coupled to the pore-gate domain (PGD) in diverse ways suitable for its specific function in each physiological context, including action potential generation, muscle contraction and relaxation, hormone and neurotransmitter secretion, and cardiac pacemaking. However, some VSD-containing proteins lack a PGD. Voltage-sensing phosphatase contains a cytoplasmic phosphoinositide phosphatase with similarity to phosphatase and tensin homolog (PTEN). Hv1, a voltage-gated proton channel, also lacks a PGD. Within Hv1, the VSD operates as a voltage sensor, gate, and pore for both proton sensing and permeation. Hv1 has a C-terminal coiled coil that mediates dimerization for cooperative gating. Recent progress in the structural biology of VGICs and VSD proteins provides insights into the principles of VSD coupling conserved among these proteins as well as the hierarchy of protein organization for voltage-evoked cell signaling. Keywords: voltage sensor, membrane potential, phosphoinositide, proton, gating Membrane excitation plays fundamental roles in these ion channels were identified through studies the functions of neurons, muscle, endocrine cells, and involving biochemistry, molecular genetics,4) and D D 5),6) electric organs. Na and K conductances through molecular cloning. Proteins comprising Nav,Kv, D the membrane, mediated by voltage-gated Na and and Cav channels share a common architecture that D K ion (Nav and Kv) channels, respectively, were includes a voltage sensor domain (VSD) consisting of the first elements shown to underlie the membrane four transmembrane helices and a pore-gate domain excitability of nerves.1) Later, voltage-gated Ca2D (PGD) consisting of two transmembrane helices with (Cav) channels were identified in muscle, neurons, an intervening turret region (Fig. 1). These voltage- and endocrine cells.2),3) The molecular correlates of gated ion channels (VGICs), together with related 1 D D * Department of Physiology, Graduate School of Medicine, Kcv: virus-derived K channel; Kir: inward rectifier K ;Kv: Osaka University, Suita, Japan. voltage-gated KD; MPO: myeloperoxidase; NADPH: nicotinamide 2 D * Graduate School of Frontier Bioscience, Osaka adenine dinucleotide phosphate; Nav: voltage-gated Na ; NHE: University, Suita, Japan. sodium proton exchanger; PBM: phosphoinositide-binding † Correspondence should be addressed: Y. Okamura, motif; PD: phosphatase domain; PGD: pore-gate domain; Department of Physiology, Graduate School of Medicine and PHD: pleckstrin homology domain; PI: phosphoinositide; PS: Graduate School of Frontier Bioscience, Osaka University, Suita, pregnenolone sulfate; PtdIns(3,4)P2: phosphatidylinositol-3,4- Osaka 565-0871, Japan (e-mail: [email protected]. bisphosphate; PtdIns(3,4,5)P3: phosphatidylinositol-3,4,5-tri- ac.jp). sphosphate; PtdIns(3,5)P2: phosphatidylinositol-3,5-bisphosphate; Abbreviations: Anap: 3-(6-acetylnaphthalen-2-ylamino)- PtdIns(4,5)P2: phosphatidylinositol-4,5-bisphosphate; PtdIns4P: 2-aminopropanoic acid; CatSper: cation channel of sperm; Cav: phosphatidylinositol-4-monophosphate; PTEN: phosphatase and voltage-gated Ca2D; CNG: cyclic nucleotide-gated; cryo-EM: tensin homolog (deleted on chromosome ten); ROS: reactive cryoelectron microscopy; EPR: electron paramagnetic resonance; oxygen species; RyR: ryanodine receptor; SNARE complex: soluble FRET: fluorescence resonance energy transfer; GEVI: gene- NSF attachment protein receptor complex; TAPP1: tandem PH encoded voltage indicator; GFP: green fluorescent protein; H2O2: domain containing protein 1; TPC: two-pore channel; TRIC: hydrogen peroxide; HCN channel: hyperpolarization-activated, trimeric intracellular channel; TRP: transient receptor potential; cyclic nucleotide-gated channel; HOCl: hypochlorous acid; VCF: voltage clamp fluorometry; VGIC: voltage-gated ion HVCN1: hydrogen voltage-gated channel 1 (gene name for Hv1, channel; VSD: voltage sensor domain; VSLD: voltage-sensor like voltage-gated proton channel); HVRP1: Hv1-related protein 1; domain; VSP: voltage-sensing phosphatase. doi: 10.2183/pjab.95.010 ©2019 The Japan Academy 112 Y. OKAMURA and Y. OKOCHI [Vol. 95, a Voltage-sensor domain Pore-gate domain x 4 : tetramer Voltage-gated or ion channel 2 or 4 homologous repeats in single subunit Phosphatase domain C2 domain VSP x 1: monomer Coiled coil Hv1/VSOP x 2: dimer Pore-gate Voltage-sensor b domain domain S6 S4 S5 + + + + S1 + S2 S3 Fig. 1. Scheme of the membrane topologies of VGIC and two VSD-containing proteins. a. VSP and Hv1. VGIC contains the voltage- sensor domain (VSD) and the pore-gate domain (PGD). Most of VGICs contain N-terminal and C-terminal cytoplasmic domains which are important for subunit assembly and gating (not specifically illustrated here). VSP has the transmembrane VSD and the cytoplasmic enzyme region which consists of the phosphatase domain and C2 domain. There is a short linker between the VSD and PD (called VSD-PD linker) (not illustrated here). Transmembrane region of Hv1, the voltage-gated proton channel, corresponds to the VSD of VGIC and VSP. Hv1 contains the cytoplasmic coiled coil region which is critical for dimer formation and gating. b. Illustration of one unit of VSD (consisting of four transmembrane helices) and PGD of domain-swapped type VGIC. Helical linker connects the two domains. See Fig. 8 for domain-nonswapped type VGIC. proteins, such as hyperpolarization-activated, cyclic more recently, with single particle cryo-electron nucleotide-gated channels (HCN channels) and two- microscopy (cryo-EM) (Fig. 2).8) One remarkable pore channels (TPCs), are all classified as members finding from recent studies of the structures of VGICs 7) of the VGIC superfamily. In Kv and HCN channels, is that the coupling of the VSD to the PGD is diverse one subunit contains a single VSD and PGD, and and optimized for the physiological functions of four subunits assemble into one channel. In Nav and each individual class of VGIC. Moreover, in several Cav, one channel is formed by a long polypeptide atypical VSD-containing proteins, the coupling of consisting of four homologous repeats, each of which the VSD is not for driving a PGD. In voltage-sensing contains a single VSD and PGD. In TPCs, one phosphatases, for example, the VSD is coupled to channel is formed by two homologous subunits, each the enzyme and regulates phosphatase activity. In containing two repeats. Transient receptor potential the voltage-gated proton channel, Hv1, which lacks (TRP) channels are ion channels transducing multi- a PGD, the VSD operates as an ion permeation modal chemical and physical stimuli, such as stretch, pathway as well as a voltage sensor, and two VSDs temperature change, and oxygen concentration, into interact with each other for cooperative gating. How intracellular signals. The transmembrane regions of protons selectively permeate through the VSD in Hv1 TRP channels have a molecular architecture that remains a mystery. In addition, cyclic nucleotide- resembles that of VGICs, consisting of a voltage- gated (CNG) channels and many TRP channels have sensor like domain (VSLD) and a PGD. immobile VSDs. The protein structures of most VGIC species In this article, we first summarize the structure– have been solved using X-ray crystallography and, function relationship and molecular mechanisms of No. 3] Principles and diversity of voltage sensor domain proteins 113 Kv1.2-Kv2.1 chimera NavPaS TPC1 Cav1.1 HCN1 Slo1 Ci-VSP mHv1cc Fig. 2. Gallery of protein structures of VGICs and VSP (taken from ref. 157). Kv1.2–Kv2.1 chimera: human voltage-gated potassium channel (PDB ID: 2R9R), TPC1: plant two-pore cation channel (PDB ID: 5E1J), Cav1.1: human skeletal muscle-type voltage-gated calcium channel (PDB ID: 5GJV), NavPaS: insect voltage-gated sodium channel (PDB ID: 5X0M), HCN1: human hyperpolarization- activated cyclic nucleotide-gated channel (PDB ID: 5U6O), Slo1: large conductance calcium-activated voltage-gated potassium channel from Aplysia (PDB ID: 5TJ6), Ci-VSP: sea squirt voltage-sensing phosphatase (full length model based on coordinates from PDB ID: 3AWF and PDB ID: 4G7V), mHv1cc: mouse voltage-gated proton channel in a chimeric form containing part of S2–S3 from Ci-VSP and the coiled coil structure of GCN4 (dimer model based on coordinates from PDB ID: 3WKV). NavPaS, Kv1.2–Kv2.1 chimera and Slo1 have four VSDs within a tetramer. TPC1 has four VSDs within a dimer. Ci-VSP has a single VSD. mHv1cc has two VSDs within a dimer. coupling between the VSD and PGD in VGICs. sensing membrane voltage and is highly conserved Then, we describe VSD-effector coupling and VSD- among VGICs. It consists of four transmembrane VSD coupling in two VSD-containing proteins, helices, S1–S4, with S4 showing a signature pattern voltage-sensing phosphatase (VSP) and Hv1, respec- of amino acid alignment: positively charged residues tively, both of which do not contain a canonical aligned with two intervening hydrophobic residues PGD. We also state more emerging functions of the (Fig. 3a).9) The positive charges in S4 are counter- VSD-like region in other proteins and the hierarchy balanced by multiple negative charges on acidic of voltage signals.
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